Volume 70, Issue 10

Plants are the primary source of nutrition and essential to maintain life on earth. They have evolved very delicate and advanced photo-sensory antennae to sense their outer environment and transduce the received information for their growth and development accordingly. This “light switch” phenomenon of plants has slowly being unraveled and various plant photoreceptors, their role in downstream molecular signaling, mutual interaction, response to circadian cycle and light signals have been discovered. The photosensory antennae in plants; phytochromes, cryptochromes and phototropins play a very crucial role in sensing the ambient light intensities. By direct interaction with the environment through these photosensory antennae, plants shift their homeostasis to regulate their growth and development. The phytochrome light receptors of plants are responsive to R/FR light and by inducing signaling pathways, trigger the physiological responses such as germination and flowering. The phytochromes also directly contribute to plant development by affecting its photosynthetic rate. To elucidate the role of phytochromes in plant metabolism, this review will focus on the importance of phytochromes, their mechanism of action and their application as an emerging field in plant biology.

The α-amylase enzyme specificity has been classified in the Carbohydrate-Active enZyme (CAZy) database into the families GH13, GH57, GH119 and eventually also GH126. α-Amylase is a glycoside hydrolase (GH) that catalyses in an endo-fashion the hydrolysis of the α-1,4-glucosidic linkages in starch and related α-glucans employing the retaining reaction mechanism. The family GH13 is the main α-amylase family with more than 28,000 members and 30 different specificities. The entire family GH13 has already been divided into 40 subfamilies; the α-amylase enzyme specificity being found in the subfamilies GH13 1, 5, 6, 7, 15, 19, 24, 27, 28, 32, 36 and 37. The present in silico study delivers a proposal to create a novel GH13 subfamily with the specificity of α-amylase. The proposal is based on a detailed bioinformatics analysis consisting of sequence, structural and evolutionary comparison of experimentally characterized α-amylases from, e.g., Bacillus aquimaris, Anoxybacillus sp. SK3-4 and DT3-1 and Geobacillus thermoleovorans, and hypothetical proteins, accompanied by α-amylases from well-established GH13 subfamilies and by closely related amylolytic enzymes (mainly from the subfamily GH13 31). Three sequence-structural features can be ascribed to the members of the newly proposed GH13 subfamily: (i) the pair of adjacent tryptophan residues positioned between the CSR-V and CSR-II in the helix α3 of the catalytic TIM-barrel; (ii) the sequence LPDlx in their CSR-V; and (iii) a ~30-residue long C-terminal region with a motif of five conserved aromatic residues. From the evolutionary point of view, the novel GH13 α-amylase subfamily is most closely related to fungal and yeast α-amylases classified in the subfamily GH13_1.

The cell-free extract of Bacillus safensis LAU 13 strain (GenBank accession No: KJ461434) was used for green biosynthesis of silver nanoparticles (Ag-NP s ). Characterization of Ag-NP s was carried out using UV-VIS spectroscopy, Fourier-transform infrared spectroscopy, and transmission electron microscopy. Evaluation of synthesized Ag-NPs as antimicrobial agents was done using multi-drug resistant clinical isolates as well as their synergistic effects when combined with some selected antibiotics. Furthermore, potential of Ag-NP s as antimicrobial additives in paint was demonstrated. The Ag-NP s have maximum absorbance at 419 nm, with peaks at 3308, 2359, 1636, and 422 cm −1 , indicating that proteins were the capping and stabilisation molecules in the synthesis of Ag-NP s . The particles were spherical shaped having size of 5-95 nm, with silver as the prominent metal from the energy dispersive X-ray analysis, while selected area electron diffraction pattern agrees well with the crystalline nature and face-centred cubic phase of Ag-NPs. Inhibition of Staphylococcus aureus, Escherichia coli, Klebsiella granulomatis and Pseudomonas aeruginosa was achieved at 100 μg/mL. Improvement of activities of augmentin, ofloxacin and cefixime to the tune of 7.4-142.9% was achieved in synergistic study, while total inhibitions of P. aeruginosa, S. aureus, Aspergillus flavus and Aspergillus fumigatus were achieved in Ag-NP s -paint admixture. The Ag-NP s showed potent antioxidant and larvicidal activities with IC 50 and LC 50 of 15.99 and 42.19 μg/mL, respectively. The present study demonstrated that the biosynthesized Ag-NPs have potent biological activities, which can find applications in diverse areas. The report adds to the growing biotechnological relevance of B. safensis.

In this study, both morphological characters and molecular techniques were used to determine the taxonomic position of the local endemic Agropyron deweyi within the genus. Plant length, inflorescence type, flower, leaf, spike, spikelet, indumentum and lemma structures were studied as morphological characters. DNA sequences of the nuclear ribosomal DNA internal transcribed spacer region (nrDNA ITS) and four chloroplast DNA regions [trnT-trnL intergenic spacer (IGS), trnL intron, trnL-trnF IGS and matK gene] were analyzed as part of molecular investigations. The results demonstrated that no remarkable morphological differences exist among studied specimens of A. deweyi and A. cristatum complex. Based on 12 sequences of 4 Agropyron taxa produced from the current study and different numbers of sequences obtained from GenBank, our results support the conclusion that the taxonomic status of Agropyron deweyi is synonymous to that of the species A. cristatum. Sequence divergences were estimated using Kimura two-parameter model, and the phylogenetic analyses were performed using the maximum parsimony (MP) and maximum likelihood (ML) methods. No genetic variation was observed among samples of native A. deweyi and A. cristatum complex regardless of the analyzed region. Agropyron deweyi was especially very similar to A. cristatum subsp. pectinatum var. pectinatum in terms of morphological characters such as inflorescence rachis tough at maturity, spikelets with distinct gaps, indumentum glabrous, and lemma glabrous. Therefore, when all morphological and molecular results are taken into account, A. deweyi and A. cristatum subsp. pectinatum var. pectinatum should be considered as synonym.

Genetic structure of the mountain dwarf pine (Pinus mugo) and Scots pine (P. sylvestris) natural populations along with their 4 putative hybrid swarms in Slovakia were analyzed using 12 enzyme loci. The study aimed in evaluation of postulated hybrid status of the swarms on molecular level. Based on semidiagnostic allele frequencies of the MDH B, MDH C, 6PGDH B, ADH 1 and F-EST loci in their megagametophytes and vegetative buds, the conspicuous genetic differences were detected between P. mugo and P. sylvestris populations. The putative hybrid swarms seem to incline by their genetic structure to P. mugo rather than to P. sylvestris. Obtained results indicate independent developmental history of each of the scored hybrid swarms. The putative hybrid swarm in Sucha Hora was found to be a mixed stand consisting prevailingly of pure-species individuals of P. mugo and P. sylvestris. On the contrary, the putative hybrid swarms in Habovka, Tisovnica and Terchova are supposed to be of hybrid origin.

The present study aimed to reveal genetic diversity and population structure in three Onosma species i.e., Onosma bulbotrichum DC., O. elwendicum Wettst. and O. sericeum Willd. (Boraginaceae) growing in Iran. This study was performed to check if the ISSR molecular markers could be used in the Onosma species delimitation. We also investigated the infra-specific morphological variability and if we can identify infra-species taxonomic forms. The present study revealed within species genetic and morphological diversity. ISSR data could delimit the studied species as they were separated from each other in NJ tree, maximum parsimony and STRUCTURE analysis. AMOVA and Hickory tests produced significant genetic difference among the studied species. The Mantel test showed no correlation between genetic distance and geographical distance of the populations for all three species. Reticulation analysis and Nm estimation showed gene flow among these populations. We could identify a few ISSR loci that are adaptive. Data obtained can be used in conservation of these medicinal plants in Iran.

Salinity stress is one of the most serious environmental factors limiting plant growth and productivity in large areas around the world. Priming approach was adopted to study the effect of glycinebetaine (GB) on enhancing salt tolerance of sensitive wheat cultivar (Gomeza 7). The caryopsis were primed in different concentrations of GB (25, 50, 100 mM) for 24 h, and then treated with or without 150 mM NaCl added to 1/4-modified Hoagland solution (MHS). The NaCl treatment lasted 38 d under natural environmental conditions. Salt stress reduced all growth parameters measured: fresh mass, dry mass, relative growth rate, for the shoots and roots, and relative water content (RWC). Salt imposition increased the level of Na + and Cl − , and reduced Ca +2 and K + levels in both shoots and roots. Exogenous application of GB alleviated the deleterious effects of salinity on growth and mineral contents, the effect was more pronounced with 25 mM GB. Priming of caryopsis in GB counteracted the increase in the plasma membrane (PM) permeability and increased the cell solute potential, which was decreased by salinity. GB priming also increased the GB and glutathione concentrations, and reduced proline (Pro) as well as lipid peroxidation. Salt tolerance enhancement by GB priming might be occurred through reduced lipid peroxidation, increased GB and glutathione resulting in PM protection, and eventually ion homeostasis. The study is a valuable confirmation for enhancing salt adaptation via GB priming, which is of general interest for agriculture practices.

As one of the essential macroelements, nitrogen (N) plays an important role in plant growth and development. In order to know the effect of different N levels on the rice plant growth and carbon-nitrogen metabolism, we analyzed the rice growth phenotype, leaf SPAD value, photosynthesis, carbon-nitrogen metabolic status and gene expression profile under four different N levels (0×N, 0.1×N, 1×N and 5×N). The plant height and dry weight increased with increasing N levels, whereas an opposite trend was observed for the root length, which decreased with increasing N levels. The leaf SPAD value, stem nitrate concentration, soluble proteins, photosynthetic rate, stomatal conductance and total nitrogen concentration increased with increasing N levels, whereas an opposite trend was observed for soluble carbohydrates and carbon/nitrogen ratio which decreased with increasing N levels. Metabolite profile analysis revealed that the low N treatment caused visible decreases in the concentrations of total sugars and organic acids in the leaves, while caused visible increases in the concentrations of total sugars, organic acids and free amino acids in the roots. Gene expression analysis showed that the transcriptional levels of 5 genes (GS1;3, NADH-GOGAT1, NADH-GOGAT2, PEPC4 and PEPC7) altered significantly under four different N levels.

The lack of an efficient transformation system for Pinellia ternata is a major bottleneck in the functional annotation of genes and development of increased stress resistance of this medicinal herb. Here we describe the efficient transformation of P. ternata using petioles from submerged cultures as explants.We used the Agrobacterium strain EHA105, which harbored a pCAMBIA2300-35S-GUS-CaMVterm plasmid containing β-glucuronidase (GUS) as the reporter gene and nptII as the selectable marker. The recovery of transgenic plants was achieved by callus induction on the selection medium followed by shoot induction on the regeneration medium under selection pressure. Pre-culture of explants and ultrasonic treatment during inoculation enhanced the transformation efficiency. Transformation frequency reached 19% based on a GUS assay of independently derived, putative transgenic lines. This transformation system should facilitate the functional characterization of genes of interest and genetic advancements in P. ternata.

In vitro shoot tips of Serbian autochthonous plums ‘Sitnica’ (Prunus domestica L.) and ‘Crvena Ranka’ (Prunus insititia L.) were tested for recovery after cryopreservation using the droplet-vitrification technique. After 30-min loading with 1.9 M glycerol and 0.5 M sucrose, explants were dehydrated at room temperature for 10, 20, 30, 40 and 50 min with PVS A3 solution (37.5% glycerol, 15% dimethylsulfoxide, 15% ethylene glycol and 22.5% sucrose) or for 60, 90 and 120 min using PVS3 solution (50% glycerol and 50% sucrose). Rewarming was performed in unloading solution containing 0.8 M sucrose for 30 or 60 min. Duration of PVS3 treatment significantly affected survival (27.3-72.7%) and regrowth (0-18.2%) of cryopreserved explants in plum ‘Sitnica’, with the highest values of both parameters being achieved with the 90-min treatment. Also, survival of explants dehydrated with PVS A3 solution significantly varied between 18.2-73.9%, depending on duration of both dehydration and unloading treatments. However, cryopreserved explants displayed a very low regrowth capacity, the highest being 10% for 30-min dehydration in combination with 60-min unloading. ‘Crvena Ranka’ exhibited a higher regrowth capacity after cryopreservation. Duration of PVS3 treatment significantly affected survival (22.2-77.8%) and regrowth (0-30.0%) of cryopreserved explants, with the highest values of both parameters being achieved with the shortest treatment duration. As regards PVS A3 treatments, both survival and regrowth significantly varied between 27.3- 81.8%, and 0-36.4%, respectively. The highest regrowth was achieved with 20- and 30-min treatment durations combined with 30-min unloading. Further optimization of the protocol is necessary to improve recovery after cryopreservation.

Natural environments tend to be variable resulting in alternating periods of high and low food availability. Therefore, animals have to be able to accommodate to sudden environmental changes by adjusting their physiology and behaviour to new conditions. We investigated how simulated food variability affects life history traits (asexual reproduction and stress tolerance) and response to environmental change in laboratory experiments with green hydra (Hydra viridissima). We assigned hydra into four groups differing in feeding frequency (high or low) and food regularity (random or stable). After 21 days of accommodation, feeding frequency was changed (increased or decreased) in half of each group, the other half was kept as a control group. Hydra showed a delayed response to environmental change (increased or decreased feeding frequency). This delay in response was greater under an unpredictable feeding scheme. Animals on a random scheme had lower budding rates and lower stress tolerance. Follow-up experiments suggest that this might be due to receiving food on subsequent days, since we found that animals fed daily have lower budding rates than those fed on alternate days. We hypothesize that frequent feeding might cause high levels of oxidative/xenobiotic stress which could overwhelm the defence system of these animals.

Three species belonging to the family Tylenchidae, Malenchus nanellus, M. undulatus and Tylenchus naranensis, are reported for the first time for Iran. These species are characterized based on morphological and morphometric data. The Iranian population of M. nanellus is characterized by its body length ranging from 410-485 μm, cuticle annuli 1.1- 1.5 μm wide at mid-body and lateral field with two crenate lines, starting at the mid-region of procorpus and ending near 1/3 of tail length. The population of M. undulatus is characterized by its 458-526 μm body length, cuticle coarsely annulated, annuli 1.8-2.4 μm wide at mid-body, lateral field with crenate incisures, beginning at about half of the stylet length, ending at middle of tail, head narrower than adjacent body, median bulb well developed with prominent valve plates and functional males in population. Iranian population of T. naranensis, is characterized by having a 631-774 μm body length, lateral field with four lines, outer lines crenate, a stylet length ranging from 10-11 μm and a tail of 102-131 μm long. Molecular phylogenetic analyses based on partial 28S rDNA sequences of T. naranensis revealed its close affinity with the genus Filenchus. Other Tylenchidae genera, such as Aglenchus (including the newly sequenced isolate from Iran) and Coslenchus were sister taxa and formed a well-supported clade. Malenchus exiguus, a previously reported species from Iran and sequenced in the present study, formed a monophyletic clade with other species of Malenchus and Lelenchus leptosoma.

A new species belonging to genus Ctenobelba (Caucasiobelba) by Subías & Shtanchaeva, 2010 is described and a firstly recorded species of genus Papillacarus Kunst, 1959 is redescribed using SEM micrographs.

This paper presents information on the oviposition, embryonic development, egg production, and reproductive success in water mites (Hydrachnidia) under laboratory conditions. The study was carried out on four common and numerous species in Poland: Hydrodroma despiciens (lake population), Hygrobates fluviatilis (river population), Hygrobates longipalpis (lake and river populations) and Mideopsis orbicularis (lake and river populations). For H. longipalpis and M. orbicularis there were no significant differences between the number of eggs deposited by females from river and lake populations. The time of embryonic development in all species was relatively similar. In all analysed species, the mean time of embryonic development shortened with the lengthening of the day. All species were characterised by very high egg hatching success.

Aeolothrips iranicus sp. n. is described and illustrated from Fars Province, south of Iran. This is the twenty first species of the genus from Iran and in contrast to most species described in the genus Aeolothrips Haliday is bicolored. The status of the new species in comparison to apparently closely related species including A. collaris Priesner, A. pulcher Oettingen, A. tauricus Derbeneva and A. montivagus Priesner is discussed. Moreover, it is shown that the spermatheca of A. iranicus and A. collaris are different.

We investigated the influence of a colony of great cormorants on the skull morphometry of yellow-necked mice (Apodemus flavicollis) and bank voles (Myodes glareolus) of three age groups trapped in the territory of the colony in 2011- 2014. In general, most of the skull differences in both species were related to character length (skulls tended to become longer). In A. flavicollis, the skull size differences between zones were most expressed in subadult and adult individuals, while in M. glareolus they were most expressed in juveniles, with only a few characters different in adults and none in subadult voles. For both species, the largest skull characters were found mostly in the zone characterized by both the greatest number of cormorant nests and the longest and strongest influence by the colony. Concluding we confirm that the great cormorant colony has an influence on the skull morphometry of A. flavicollis and M. glareolus and we hypothesize that these differences enhance the ability of survival in specific conditions.

Increased risk of osteoporosis in patients with diabetes mellitus may be related to hyperglycemia. However, the potential mechanisms accounting for diabetic bone disorder remain unresolved. The present study investigated the effects of high glucose-associated osmolality on differentiation of primary rat calvarial osteoblasts. Osteoblastogenic differentiation was determined by bone nodule staining for mineralization assay, enzyme-linked immunosorbent assay for type I collagen production and real-time polymerase chain reaction (PCR) for osteoblastogenic marker gene expression. Adipocytogenic differentiation was assessed by oil red O staining for lipid accumulation and real-time PCR for adipocytogenic marker gene expression. The phosphorylations of protein kinase A (PKA) and Akt were measured with or without specific inhibitors to confirm osmolality involved signalling pathways. The results showed that high glucose-associated osmolality significantly promoted adipocytogenic differentiation, manifested by increased lipid droplet formation and gene expression of adipocytogenic markers including adipocyte fatty acid binding protein (aP2), adipsin and peroxisome proliferator-activated receptor gamma (PPARγ). Meanwhile, high glucose-associated osmolality inhibited osteoblastogenic differentiation, characterized by decreased collagen I protein production and cell mineralization, as well as gene expression of osteoblastogenic markers including collagen I, osteocalcin and runt-related transcription factor 2 (Runx2). More importantly, we demonstrated for the first time that high glucose-associated osmolality induced adipocytogenic differentiation and suppressed osteoblastogenic differentiation in a PKA and phosphatidylinositol 3-kinase (PI3K)/Akt-dependent manner. These results indicated that osmolality was involved in high glucose-induced osteoblast trans-differentiation into adipocyte-like cell and suppression of cellular osmolality could provide novel therapeutic approach for diabetic osteopenia.

Rooibos is widely used in folk medicine, but its influence on reproductive processes has been studied insufficiently. The aim of present study was to examine possible involvement of rooibos on the release of progesterone (P4), testosterone (T) and insulin-like growth factor I (IGF-I), as well as its response to gonadotropin - follicle-stimulating and luteinizing hormone (FSH+LH), the most known hormonal regulators for reproduction. Rabbit ovarian fragments were cultured in the presence of rooibos (1, 10 and 100 μg/mL medium), FSH+LH (0.01, 0.1 and 1 IU/mL medium) and rooibos (10 μg/mL medium) in combination with FSH+LH (0.01, 0.1 and 1 IU/mL medium). The release of hormones was evaluated by using radioimmunoassay. It was observed that rooibos addition to culture medium stimulated P4 and inhibited T release. IGF-I release was significantly stimulated by rooibos at all doses added. FSH+LH when given alone promoted IGF-I, but not P4 or T release. The addition of rooibos induced the suppressive effect of FSH+LH on P4 release, the stimulatory action of FSH+LH on T output and reduced the response of IGF-I to FSH+LH treatment. These data suggest that rooibos is able to directly affect the release of steroid (P4 and T) and peptide (IGF-I) hormones by rabbit ovarian cells and to modulate their response to gonadotropins.

Raykov et al. (2012) recorded the European mudminnow (Umbra krameri) from the Black Sea, at a depth of 36.3-41 m. Morphometric comparison of the pictured specimen with 10 adult U. krameri and published data was conducted which excluded its taxonomic affiliation to Umbridae family.